Introductio2.edited (2)

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Introduction Prevalence of hypertension worldwide and its impact on public health 1. Hypertension is a common concern in today's society, with increasing incidence due to unhealthy habits. 2. Reports suggest that the prevalence of hypertension worldwide is as high as 10-20%. 3. By 2025, the number of hypertensive patients is expected to exceed 1.5 billion. 4. Hypertension is a major risk factor for cardiovascular disease, stroke, and other health problems. The prevalence of hypertension worldwide and its impact on public health emphasizes the increasing incidence of hypertension due to unhealthy habits and the significant impact it has on public health. Reports suggest that the prevalence of hypertension worldwide is high and is expected to increase in the future. Hypertension is a major risk factor for cardiovascular disease, stroke, and other health problems. The potential of functional food ingredients to help manage hypertension 1. Functional food ingredients are those that provide health benefits beyond basic nutrition. 2. Peptides derived from food proteins have been shown to have ACE inhibitory activity, which can help lower blood pressure. 3. Walnut dregs are a by-product of walnut production that contains valuable proteins and peptides. 4. This work explores the potential of walnut hydrolysates as functional food ingredients with blood- pressure-lowering effects. This focuses on the potential of functional food ingredients to help manage hypertension. Peptides derived from food proteins have been shown to have ACE inhibitory activity, which can help lower blood pressure. Walnut dregs are a by-product of walnut production that contains valuable proteins and peptides. This work explores the potential of walnut hydrolysates as functional food ingredients with blood-pressure-lowering effects. By exploring these subtopics, the document aims to provide insights into the potential of functional food ingredients to help manage hypertension and improve public health. Chapter 2: Preparation and Identification of Walnut Hydrolysate

Enzymatic hydrolysis of walnut dregs 1. Enzymatic hydrolysis is an effective method for preparing bioactive peptides from food proteins. 2. Walnut dregs were used as a source of proteins and peptides for hydrolysis. 3. The hydrolysis was carried out using protease enzymes, which were added to the walnut dregs. 4. The hydrolysis process was optimized to achieve maximum ACE inhibitory activity. Enzymatic hydrolysis is a widely used method for preparing bioactive peptides from food proteins. In this chapter, walnut dregs were used as a source of proteins and peptides for hydrolysis. The hydrolysis was carried out using protease enzymes, which were added to the walnut dregs. The hydrolysis process was optimized to achieve maximum ACE inhibitory activity. The results showed that the optimal conditions for hydrolysis were a substrate concentration of 5%, an enzyme concentration of 2%, and a hydrolysis time of 4 hours. These conditions resulted in a hydrolysate with high ACE inhibitory activity. Subtopic 2: Purification of ACE inhibitory peptides 1. The hydrolysate was subjected to ultrafiltration to separate the peptides based on their molecular weight. 2. The peptides with high ACE inhibitory activity were further purified using reverse-phase high- performance liquid chromatography (RP-HPLC). 3. The purified peptides were identified using mass spectrometry. 4. The identified peptides were found to be mainly composed of dipeptides and tripeptides. After enzymatic hydrolysis, the hydrolysate was subjected to ultrafiltration to separate the peptides based on their molecular weight. The peptides with high ACE inhibitory activity were further purified using RP-HPLC. The purified peptides were identified using mass spectrometry. The identified peptides were found to be mainly composed of dipeptides and tripeptides. The results suggest that these peptides may have the potential as functional food ingredients with blood-pressure-lowering effects. Subtopic 3: Characterization of ACE inhibitory peptides 1. The ACE inhibitory activity of the purified peptides was evaluated using in vitro assays.

2. The peptides were found to have high ACE inhibitory activity, with IC50 values ranging from 0.05 to 0.15 mg/mL. 3. The peptides were also found to have antioxidant activity, which may contribute to their health benefits. 4. The amino acid sequences of the identified peptides were analyzed to determine their structure and potential bioactivity. The ACE inhibitory activity of the purified peptides was evaluated using in vitro assays. The peptides were found to have high ACE inhibitory activity, with IC50 values ranging from 0.05 to 0.15 mg/mL. The peptides were also found to have antioxidant activity, which may contribute to their health benefits. The amino acid sequences of the identified peptides were analyzed to determine their structure and potential bioactivity. The results showed that the identified peptides were mainly composed of hydrophobic amino acids, which are known to contribute to ACE inhibitory activity. The peptides were also found to contain amino acids with potential bioactivity, such as arginine and proline. Subtopic 4: continuation 1. Enzymatic hydrolysis of walnut dregs can produce peptides with high ACE inhibitory activity. 2. The purified peptides were found to have the potential as functional food ingredients with blood- pressure-lowering effects. 3. The identified peptides were mainly composed of dipeptides and tripeptides with hydrophobic amino acids and potential bioactivity. 4. Further research is needed to explore the potential health benefits of these peptides and their applications in functional foods. In conclusion, enzymatic hydrolysis of walnut dregs can produce peptides with high ACE inhibitory activity. The purified peptides were found to have the potential as functional food ingredients with blood-pressure-lowering effects. The identified peptides were mainly composed of dipeptides and tripeptides with hydrophobic amino acids and potential bioactivity. Further research is needed to explore the potential health benefits of these peptides and their applications in functional foods. Overall, this chapter provides valuable insights into the preparation and identification of walnut hydrolysates and their potential as functional food ingredients. Chapter 3: Antihypertensive Activity of Walnut Hydrolysate

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Subtopic 1: Changes in blood pressure and body weight of SHR 1. Spontaneously hypertensive rats (SHR) were used as a model to evaluate the antihypertensive activity of walnut hydrolysate. 2. The rats were administered with different doses of walnut hydrolysate for 4 weeks. 3. The results showed that the hydrolysate had a dose-dependent antihypertensive effect, with the highest dose (500 mg/kg) showing the most significant effect. 4. The hydrolysate also had no significant effect on the body weight of the rats. 5. These results suggest that walnut hydrolysate has potential as a natural antihypertensive agent. Spontaneously hypertensive rats (SHR) were used as a model to evaluate the antihypertensive activity of walnut hydrolysate. The rats were administered with different doses of walnut hydrolysate for 4 weeks. The results showed that the hydrolysate had a dose-dependent antihypertensive effect, with the highest dose (500 mg/kg) showing the most significant effect. The hydrolysate also had no significant effect on the body weight of the rats. These results suggest that walnut hydrolysate has potential as a natural antihypertensive agent. The findings of this study provide valuable insights into the potential health benefits of walnut hydrolysate and its applications in managing hypertension. Subtopic 2: Changes in serum biochemical parameters of SHR 1. The serum biochemical parameters of the rats were evaluated to determine the effect of walnut hydrolysate on their health. 2. The results showed that the hydrolysate had no significant effect on the levels of total cholesterol, triglycerides, or glucose. 3. The hydrolysate also had no significant effect on the levels of liver and kidney function markers. 4. These results suggest that walnut hydrolysate is safe for consumption and does not have any adverse effects on the health of the rats. 5. Further research is needed to determine the long-term safety and efficacy of walnut hydrolysate in humans. The serum biochemical parameters of the rats were evaluated to determine the effect of walnut hydrolysate on their health. The results showed that the hydrolysate had no significant effect on the

levels of total cholesterol, triglycerides, or glucose. The hydrolysate also had no significant effect on the levels of liver and kidney function markers. These results suggest that walnut hydrolysate is safe for consumption and does not have any adverse effects on the health of the rats. Further research is needed to determine the long-term safety and efficacy of walnut hydrolysate in humans. The findings of this study provide important information on the safety and potential health benefits of walnut hydrolysate, which can be used to develop functional foods and nutraceuticals for managing hypertension. Subtopic 3: Mechanisms of antihypertensive activity of walnut hydrolysate 1. The mechanisms of the antihypertensive activity of walnut hydrolysate were investigated. 2. The hydrolysate was found to inhibit the activity of angiotensin-converting enzyme (ACE), which is involved in the regulation of blood pressure. 3. The hydrolysate was also found to increase the activity of superoxide dismutase (SOD), which is an antioxidant enzyme that protects against oxidative stress. 4. These results suggest that the antihypertensive activity of walnut hydrolysate may be due to its inhibition of ACE and its antioxidant activity. 5. Further research is needed to determine the specific bioactive peptides in walnut hydrolysate that are responsible for its antihypertensive activity. The mechanisms of the antihypertensive activity of walnut hydrolysate were investigated. The hydrolysate was found to inhibit the activity of angiotensin-converting enzyme (ACE), which is involved in the regulation of blood pressure. The hydrolysate was also found to increase the activity of superoxide dismutase (SOD), which is an antioxidant enzyme that protects against oxidative stress. These results suggest that the antihypertensive activity of walnut hydrolysate may be due to its inhibition of ACE and its antioxidant activity. Further research is needed to determine the specific bioactive peptides in walnut hydrolysate that are responsible for its antihypertensive activity. The findings of this study provide important insights into the potential mechanisms of action of walnut hydrolysate and its potential as a natural antihypertensive agent. Chapter 4: Effect of Walnut Hydrolysate on Gut Microbiota in SHR Subtopic 1: Materials and Methods 1. The study used SHR as a model to investigate the effect of walnut hydrolysate on gut microbiota.

2. The rats were divided into three groups: control, low-dose, and high-dose walnut hydrolysate. 3. The gut microbiota of the rats was analyzed using 16S rDNA amplicon sequencing. 4. The diversity and composition of the gut microbiota were evaluated using alpha and beta diversity analysis. 5. The relative abundance of specific bacterial taxa was also analyzed. The study used SHR as a model to investigate the effect of walnut hydrolysate on gut microbiota. The rats were divided into three groups: control, low-dose, and high-dose walnut hydrolysate. The gut microbiota of the rats was analyzed using 16S rDNA amplicon sequencing. The diversity and composition of the gut microbiota were evaluated using alpha and beta diversity analysis. The relative abundance of specific bacterial taxa was also analyzed. This study provides valuable information on the methodology used to investigate the effect of walnut hydrolysate on gut microbiota. Subtopic 2: Effect of walnut hydrolysate on gut microbiota diversity and composition 1. The results showed that walnut hydrolysate had no significant effect on the alpha diversity of gut microbiota. 2. However, the beta diversity analysis showed that the gut microbiota of the high-dose group was significantly different from that of the control group. 3. The relative abundance of specific bacterial taxa was also affected by walnut hydrolysate. 4. The high-dose group had a higher relative abundance of beneficial bacteria, such as Lactobacillus and Bifidobacterium. 5. The low-dose group had a higher relative abundance of potentially harmful bacteria, such as Desulfovibrio and Enterococcus. The results showed that walnut hydrolysate had no significant effect on the alpha diversity of gut microbiota. However, the beta diversity analysis showed that the gut microbiota of the high-dose group was significantly different from that of the control group. The relative abundance of specific bacterial taxa was also affected by walnut hydrolysate. The high-dose group had a higher relative abundance of beneficial bacteria, such as Lactobacillus and Bifid bacterium. The low-dose group had a higher relative abundance of potentially harmful bacteria, such as Desulfovibrio. Subtopic 3: Mechanisms of the effect of walnut hydrolysate on gut microbiota

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1. The mechanisms underlying the effect of walnut hydrolysate on gut microbiota were investigated. 2. The hydrolysate was found to increase the relative abundance of beneficial bacteria, such as Lactobacillus and Bifidobacterium. 3. The hydrolysate was also found to decrease the relative abundance of potentially harmful bacteria, such as Desulfovibrio and Enterococcus. 4. These results suggest that the effect of walnut hydrolysate on gut microbiota may be due to its prebiotic activity. 5. Further research is needed to determine the specific peptides in walnut hydrolysate that are responsible for its prebiotic activity. The mechanisms underlying the effect of walnut hydrolysate on gut microbiota were investigated. The hydrolysate was found to increase the relative abundance of beneficial bacteria, such as Lactobacillus and Bifid bacterium. The hydrolysate was also found to decrease the relative abundance of potentially harmful bacteria, such as Desulfovibrio and Enterococcus. These results suggest that the effect of walnut hydrolysate on gut microbiota may be due to its prebiotic activity. Further research is needed to determine the specific peptides in walnut hydrolysate that are responsible for its prebiotic activity. The findings of this study provide important insights into the potential mechanisms of action of walnut hydrolysate on gut microbiota and its potential as a functional food ingredient. Chapter 5: Walnut Hydrolysate Biscuits Subtopic 1: Formulation and optimization of walnut hydrolysate biscuits 1. The study used low-gluten flour, walnut hydrolysate, butter, sugar powder, and egg liquid as raw materials to prepare functional biscuits. 2. Single factor and orthogonal optimization experiments were conducted to evaluate the sensory scores of the biscuits and optimize the production process. 3. The optimal formula for the biscuits was found to be 45.7% flour, 11.4% walnut hydrolysate, 22.9% butter, 11.4% sugar powder, and 8.6% egg liquid. 4. The biscuits made with this formula had a unique aroma of walnut protein, a crispy texture, and comprehensive nutrition. 5. The biscuits were found to be suitable for the general public's taste and had potential as functional food products.

The study used low-gluten flour, walnut hydrolysate, butter, sugar powder, and egg liquid as raw materials to prepare functional biscuits. Single factor and orthogonal optimization experiments were conducted to evaluate the sensory scores of the biscuits and optimize the production process. The optimal formula for the biscuits was found to be 45.7% flour, 11.4% walnut hydrolysate, 22.9% butter, 11.4% sugar powder, and 8.6% egg liquid. The biscuits made with this formula had a unique aroma of walnut protein, a crispy texture, and comprehensive nutrition. The biscuits were found to be suitable for the general public's taste and had potential as functional food products. This study provides valuable information on the formulation and optimization of walnut hydrolysate biscuits. Subtopic 2: Physicochemical properties of walnut hydrolysate biscuits 1. The physicochemical properties of the biscuits were evaluated, including moisture content, water activity, and texture. 2. The moisture content of the biscuits was found to be 3.5%, which is within the acceptable range for biscuits. 3. The water activity of the biscuits was found to be 0.3, which is low enough to prevent microbial growth and maintain the shelf life of the biscuits. 4. The texture of the biscuits was evaluated using a texture analyzer, and the results showed that the biscuits had a crispy texture. 5. The biscuits were found to have good physicochemical properties and were suitable for commercial production. The physicochemical properties of the walnut hydrolysate biscuits were evaluated. The moisture content of the biscuits was found to be 3.5%, which is within the acceptable range for biscuits. This is important because moisture content affects the texture, shelf life, and overall quality of the biscuits. The water activity of the biscuits was found to be 0.3, which is low enough to prevent microbial growth and maintain the shelf life of the biscuits. This is important because water activity is a key factor in determining the stability and safety of food products. The texture of the biscuits was evaluated using a texture analyzer, and the results showed that the biscuits had a crispy texture. Subtopic 3: Health benefits of walnut hydrolysate biscuits 1. The study investigated the potential health benefits of walnut hydrolysate biscuits. 2. The biscuits were found to have high ACE inhibitory activity, which may help to lower blood pressure.

3. The biscuits were also found to have antioxidant activity, which may help to prevent oxidative stress and related diseases. 4. The biscuits were found to be a good source of protein and dietary fibre, which may help to promote satiety and weight management. 5. The biscuits were found to be suitable for people with hypertension and may have the potential as a functional food for managing blood pressure. The study investigated the potential health benefits of walnut hydrolysate biscuits. The biscuits were found to have high ACE inhibitory activity, which may help to lower blood pressure. The biscuits were also found to have antioxidant activity, which may help to prevent oxidative stress and related diseases. The biscuits were found to be a good source of protein and dietary fibre, which may help to promote satiety and weight management. The biscuits were found to be suitable for people with hypertension and may have the potential as a functional food for managing blood pressure. This study provides important insights into the potential health benefits of walnut hydrolysate biscuits and their applications in managing hypertension and promoting overall health.

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Introductio2.edited (2)

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